The major goal of this program project is to develop drugs that may prove beneficial to Alzheimer's disease (AD) patients. Paramount to the establishment of a drug or class of drugs that may have therapeutic applications in AD is the establishment of alterations (improvements or possibly further deterioration) in the neurochemical profile while undergoing drug therapy. It is the primary focus of this Neurochemistry Core to maintain a central laboratory facility which will support the various projects outlined in the program by providing assay service for a number of hormonal, cholinergic and peptidergic neurochemical markers. The services provided will be as follows; 1) radioimmunoassay (RIA) support for the various estrogens (17beta-estradiol, 17alpha-estradiol, estrone, estriol), anterior pituitary hormones (luteinizing hormone [LH] and thyroid stimulating hormone [TSH] and the thyroid hormones T3 and T4; 2) enzyme- linked immunoabsorbant assay (ELISA) service for the neurotrophic agent nerve growth factor (NGF); 3). radiometric analysis of various markers of cholinergic function (high affinity choline uptake [HACU], acetylcholine release and choline acetyltransferase [ChAT] activity) and protein kinase C (PKC) in both neural tissue and cells from neuron-enriched cultures; 4) semiquantitative measurement of high affinity NGF receptor protein (trkA) by immunoprecipitation and electrophoretic separation in both neural tissue and cells from neuron-enriched cultures; 5) radiometric monitoring of intracellular ionized (free) calcium [Ca++]i by using calcium dye Fura2/AM and photon counting spectrofluorometric procedures; 6) immunocytochemical analysis of ChAT, cyclic AMP response element binding protein (CREB), phosphorylated form of the cyclic AMP response element binding protein (P- CREB) and mitochondrial proto-oncogene protein bcl-2 levels in neural tissue and 7) in situ end-labeling of DNA oligonucleotides in brain tissue and cultured neurons to determine the extent of apoptosis in cultured neurons when exposed to cellular insults such as serum deprivation, exposure to NMDA or beta-amyloid protein,. Thus, the Neurochemistry Core will be integral to this program project by providing analytical service for the monitoring of neurochemical status while selected drugs undergo both preliminary screening and more detailed evaluation for potential therapeutic uses in the treatment of AD and related disorders.